We developed a protocol for the rapid identification and quantitation of fungi by quantitative real-time polymerase chain reaction (qPCR) in carpet. The fungi used in this study are field isolates of Alternaria alternata, Aspergillus versicolor, Cladosporium cladosporoides, and Stachybotrys chartarum. The modified spore extraction method provided superior quality, high-molecular-weight genomic ...

Dehalococcoides species are responsible for the reductive dehalogenation of an impressive range of common, persistent environmental contaminants. These microorganisms are difficult to both obtain and grow in pure culture, and so are often studied while they exist in consortia using molecular techniques. In particular, a significant number of quantitative real-time PCR (qPCR) assays targeting De...

Although viruses of haloarchaea are the predominant predator in hypersaline ecosystem, the culture studies about halovirus-host systems are infancy. The main reason is the tradition methodology (plaque assay) for virus-host interaction depends on culturable and susceptible host. Actually, more than 90% of haloarchaea are unculturable. Therefore, it is necessary to establish an approach for dete...

Porcine diarrhea caused by viruses is a major problem of the pig farming industry and can result in substantial losses of revenue. Thus, diagnosing the infectious agents is important to prevent and control diseases in pigs. We developed novel one-step real-time quantitative RT-PCR (qPCR) assays that can detect four porcine diarrheal viruses simultaneously: porcine epidemic diarrhea virus (PEDV)...

Background: It is well documented that fetal DNA can cross the placenta and is present in peripheral maternal blood during pregnancy in human. This fetal DNA also named circulating cell free fetal DNA, has emerged as a valuable source for genetic evaluation. Compared with humans, ovine species have a different structure of placental (synepitheliochorial) with no direct contact between the troph...

Laser-assisted microdissection has mainly been used in cancer studies to excise pure cell populations from heterogeneous tissues. Cancer and normal cells selected by laser-assisted microdissection have frequently been used for mRNA expression studies usually by reverse transcriptase-quantitative polymerase chain reaction (qPCR). Recently, real time immuno-qPCR was developed as a new tool for hi...

Honey-bee colony losses are an increasing problem in Western countries. There are many different causes, including infections due to various pathogens. Molecular biology techniques have been developed to reliably detect and identify honey-bee pathogens. The most sensitive, specific and reliable is the quantitative real-time polymerase chain reaction (qPCR) methodology. This review of the litera...

Real-time quantitative PCR (qPCR) is an indispensable molecular biology tool in the post-genome era. QPCR assays have become a robust and ideal tool for several applications such as clinical diagnostics, gene expression studies, biomarker discovery, and more. Using an intact genomic DNA (gDNA) template is crucial for generating meaningful qPCR data. Therefore, assessment of the gDNA integrity i...

BACKGROUND Three species of feline haemoplasma are recognised: Mycoplasma haemofelis (Mhf), 'Candidatus Mycoplasma haemominutum' (CMhm) and 'Candidatus Mycoplasma turicensis (CMt). This study compared a reverse line blot hybridization (RLB) assay for simultaneous detection of Mhf, CMhm with three separate quantitative real-time polymerase chain reaction (qPCR) assays used for diagnosis of Mhf, ...

Diagnosing and quantifying plant-parasitic nematodes is critical for efficient nematode management. Several studies have been performed intending to demonstrate nematode quantification via real-time quantitative PCR. However, most of the studies used dilution of DNA templates to make standard curves, while few studies used samples with different nematode numbers to make the standard curve, resu...